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Original article
Newly designed 11-gene panel reveals first case of hereditary amyloidosis captured by massive parallel sequencing
  1. Zuzana Chyra Kufova1,2,3,
  2. Tereza Sevcikova1,2,
  3. Jaroslav Januska4,
  4. Petr Vojta5,
  5. Arpad Boday6,
  6. Pavla Vanickova6,
  7. Jana Filipova1,2,7,
  8. Katerina Growkova1,2,7,
  9. Tomas Jelinek1,2,7,
  10. Marian Hajduch5,
  11. Roman Hajek1,2
  1. 1 Department of Haematooncology, University Hospital Ostrava, Ostrava, Czech Republic
  2. 2 Department of Clinical Studies, Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic
  3. 3 Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic
  4. 4 Cardiocentre Podlesi, Trinec, Czech Republic
  5. 5 Faculty of Medicine and Dentistry, Institute of Molecular and Translational Medicine, Palacky University, Olomouc, Czech Republic
  6. 6 Laboratory of Molecular Biology, Department of Medical Genetics, Laboratory AGEL, Novy Jicin, Czech Republic
  7. 7 Department of Biology and Ecology, Faculty of Science, University of Ostrava, Ostrava, Czech Republic
  1. Correspondence to Zuzana Chyra Kufova, Department of Haematooncology, University Hospital Ostrava, Ostrava-Poruba 708 52, Czech Republic; zuzana.chyra{at}fno.cz

Abstract

Aims Amyloidosis is caused by deposition of abnormal protein fibrils, leading to damage of organ function. Hereditary amyloidosis represents a monogenic disease caused by germline mutations in 11 amyloidogenic precursor protein genes. One of the important but non-specific symptoms of amyloidosis is hypertrophic cardiomyopathy. Diagnostics of hereditary amyloidosis is complicated and the real cause can remain overlooked. We aimed to design hereditary amyloidosis gene panel and to introduce new next-generation sequencing (NGS) approach to investigate hereditary amyloidosis in a cohort of patients with hypertrophic cardiomyopathy of unknown significance.

Methods Design of target enrichment DNA library preparation using Haloplex Custom Kit containing 11 amyloidogenic genes was followed by MiSeq Illumina sequencing and bioinformatics identification of germline variants using tool VarScan in a cohort of 40 patients.

Results We present design of NGS panel for 11 genes (TTR, FGA, APOA1, APOA2, LYZ, GSN, CST3, PRNP, APP, B2M, ITM2B) connected to various forms of amyloidosis. We detected one mutation, which is responsible for hereditary amyloidosis. Some other single nucleotide variants are so far undescribed or rare variants or represent common polymorphisms in European population.

Conclusions We report one positive case of hereditary amyloidosis in a cohort of patients with hypertrophic cardiomyopathy of unknown significance and set up first panel for NGS in hereditary amyloidosis. This work may facilitate successful implementation of the NGS method by other researchers or clinicians and may improve the diagnostic process after validation.

  • amyloid
  • haematology
  • DNA
  • molecular genetics
  • peripheral blood

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/

https://doi.org/10.1136/jclinpath-2017-204978

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    Footnotes

    • Handling editor Mary Frances McMullin.

    • Contributors ZKC, TS and KG performed the experiments. PV performed the bioinformatical analysis. ZKC, TS, PV, JJ and AB wrote the manuscript. ZKC, TS, JJ and RH designed the study. JJ, AB and PVa provided the clinical and samples background. JF, TJ, MH and RH improved and proofread the manuscript.

    • Funding This work has been realised with the support of the IRP03_2018_2020, MH CZ-DRO-FNOs/2016 and with the support by the Ministry of Health 15-29667A project: Strengthening international cooperation in science, research and education 01211/2016/RRC and Ministry of School and Education (LO1304) and Specific university research of the Faculty of Medicine (University of Ostrava) project numbers SGS 02/LF/2014-2015, SGS 09/LF/2016-2017. The project was supported by Amgen.

    • Competing interests None declared.

    • Patient consent Obtained.

    • Ethics approval Ethics Committee of University Hospital Ostrava.

    • Provenance and peer review Not commissioned; externally peer reviewed.

    • Data sharing statement BAM files are available at the corresponding author upon request.

    • Correction notice This article has been corrected since it was published Online First. The sentence ’Specifically, the 25 nucleotide insertion was identified in PRNP gene coding region…' on page 4 has been corrected. The mutation "p.Glu62Lys" has been correct throughout the paper.